LOCUS       sequence-327810-        7217 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Lentiviral plasmid for expression of mCherry with a C-terminal CAAX 
            polybasic sequence from KRras targeting mCherry to the plasma 
            membrane..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7217)
  AUTHORS   Janska L, Anandi L, Kirchberger NC, Marinkovic ZS, Schachtner LT, 
            Guzelsoy G, Carmona-Fontaine C
  TITLE     The MEMIC is an ex vivo system to model the complexity of the tumor 
            microenvironment.
  JOURNAL   Dis Model Mech. 2021 Aug 1;14(8). pii: 271783. doi: 
            10.1242/dmm.048942. Epub 2021 Aug 18.
  PUBMED    34407185
REFERENCE   2  (bases 1 to 7217)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Dis Model 
            Mech."; date: "2021-08-1"; volume: "14(8). pii: 271783. doi"; pages:
            " 10.1242/dmm.048942. Epub 2021 Aug 18"
COMMENT     Sequence Label: pLenti-mCherry-CAAX
FEATURES             Location/Qualifiers
     source          1..7217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        13..211
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             229..409
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    456..581
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1078..1311
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1496..1540
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1838..1955
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2074..2453
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2455..2653
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             2689..3396
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2834..2852)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3102..3121
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    3500..4088
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3553..3573)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3971..3982)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4091..4107)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4092..4108)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             4617..4797
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4859..5447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4939..4958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5618..6409,6410..6478))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6241..6260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6479..6583)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6658..6677)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(6849..7217,1..11)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
       61 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      121 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      181 ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg
      241 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
      301 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
      361 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
      421 acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg
      481 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
      541 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag cgggggagaa
      601 ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa tataaattaa
      661 aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct ggcctgttag
      721 aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt cagacaggat
      781 cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg catcaaagga
      841 tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa aacaaaagta
      901 agaccaccgc acagcaagcg gccggccgct gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg cactgcgtgc
     1801 gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
     1861 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     1921 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
     1981 atccagtttg gttagtaccg ggcccgggtc gactagtgat cagtgtgagg gagtgtaaag
     2041 ctggtttcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
     2101 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     2161 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     2221 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     2281 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     2341 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     2401 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     2461 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     2521 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     2581 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     2641 tctatataag cagcgcgtct aactcgaacg ctagctgtgc gatcgtttat ggtgagcaag
     2701 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
     2761 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     2821 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     2881 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     2941 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     3001 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     3061 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     3121 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     3181 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     3241 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     3301 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     3361 cgccactcca ccggcggcat ggacgagctg tacaaggaat tcaagaaaaa gaagaagaag
     3421 tcaaaaacca agtgtgttat catgtaaaaa cgttgttgtt tggggttgaa ttactcttct
     3481 agaccgcggc gcgcccgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3541 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3601 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3661 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3721 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3781 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3841 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     3901 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3961 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     4021 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     4081 ctccccgcat cgataccgtc gacctcgatc gagacctaga aaaacatgga gcaatcacaa
     4141 gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg
     4201 aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg
     4261 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     4321 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc
     4381 agaactacac accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc
     4441 tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt
     4501 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt
     4561 ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt
     4621 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     4681 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     4741 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagca
     4801 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4861 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4921 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4981 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5041 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5101 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5161 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5221 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5281 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5341 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5401 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5461 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5521 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5581 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5641 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5701 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5761 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5821 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5881 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5941 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6001 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6061 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6121 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6181 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6241 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6301 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6361 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6421 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6481 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6541 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6601 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     6661 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     6721 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     6781 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     6841 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     6901 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     6961 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     7021 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     7081 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     7141 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     7201 ctacgtatta gtcatcg
//
